JP2010004884A5 - - Google Patents
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- JP2010004884A5 JP2010004884A5 JP2009151195A JP2009151195A JP2010004884A5 JP 2010004884 A5 JP2010004884 A5 JP 2010004884A5 JP 2009151195 A JP2009151195 A JP 2009151195A JP 2009151195 A JP2009151195 A JP 2009151195A JP 2010004884 A5 JP2010004884 A5 JP 2010004884A5
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- JP
- Japan
- Prior art keywords
- reagent
- dna
- nucleic acid
- abasic
- polyamine
- Prior art date
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Claims (7)
ａ． 前記核酸を含有する試料溶液を提供する段階；
ｂ． 少なくとも１つの脱塩基部位を伴うＤＮＡを産生するために、ＤＮＡグリコシラーゼ活性を有する酵素の存在下で前記核酸を増幅する段階；
ｃ． 脱塩基ＤＮＡの分解を促進する少なくとも１つの試薬を提供する段階であって、ここで前記試薬がポリアミンである、段階；
ｄ． 少なくとも１つの脱塩基部位を伴う前記ＤＮＡの分解をひき起こすのに適した条件において前記試料溶液をインキュベートする段階、
を含む方法。 A method for reducing carryover contamination in nucleic acid polymerase chain reaction (PCR) amplification :
a. Providing a sample solution containing the nucleic acid ;
b. Amplifying said nucleic acid in the presence of an enzyme having DNA glycosylase activity to produce DNA with at least one abasic site;
c. Providing at least one reagent that promotes degradation of abasic DNA , wherein said reagent is a polyamine ;
d. Incubating the sample solution in conditions suitable to cause degradation of the DNA with at least one abasic site;
Including methods.
核酸の増幅に有用な試薬であって、ここで前記試薬が、１つ以上のオリゴヌクレオチドプライマー、核酸ポリメラーゼ、緩衝液、塩、ならびにｄＡＴＰ、ｄＣＴＰ、ｄＧＴＰおよび１つ以上のｄＴＴＰとｄＵＴＰとを含むヌクレオシド三リン酸を含む、試薬、
少なくとも１つの脱塩基部位を伴うＤＮＡを産生するのに適切な試薬、および
脱塩基ＤＮＡの分解を促進する試薬を含み、
ここで前記少なくとも１つの脱塩基部位を伴うＤＮＡを産生するのに適切な試薬がウラシル−Ｎ−ＤＮＡグリコシラーゼ活性を有し、そして前記脱塩基ＤＮＡの分解を促進する試薬がポリアミンである、反応混合物。 A reaction mixture for preventing carryover contamination in nucleic acid polymerase chain reaction (PCR) amplification , comprising:
A reagent useful for nucleic acid amplification, wherein the reagent comprises one or more oligonucleotide primers, nucleic acid polymerase, buffer, salt, and dATP, dCTP, dGTP and one or more dTTP and dUTP A reagent comprising a nucleoside triphosphate,
A reagent suitable for producing DNA with at least one abasic site, and a reagent that promotes degradation of the abasic DNA ;
Wherein the reagent suitable for producing the DNA with the at least one abasic site has uracil-N-DNA glycosylase activity and the reagent that promotes degradation of the abasic DNA is a polyamine. .
ａ） ＤＮＡグリコシラーゼ活性を有する試薬、
ｂ） 脱塩基ＤＮＡを分解することができる試薬であって、ここで前記試薬がポリアミンである試薬、及び
ｃ） 核酸の増幅に有用な試薬であって、ここで前記試薬が１つ以上のオリゴヌクレオチドプライマー、核酸ポリメラーゼ、緩衝液、塩、ならびにｄＡＴＰ、ｄＣＴＰ、ｄＧＴＰ及び１つ以上のｄＴＴＰとｄＵＴＰとを含むヌクレオシド三リン酸を含む、試薬、
を含むキット。 A kit for carrying out a method for preventing carryover contamination in nucleic acid polymerase chain reaction (PCR) amplification ,
a) a reagent having DNA glycosylase activity;
b) a reagent capable of degrading abasic DNA , wherein said reagent is a polyamine; and
c) a reagent useful for nucleic acid amplification, wherein the reagent comprises one or more oligonucleotide primers, nucleic acid polymerase, buffer, salt, and dATP, dCTP, dGTP and one or more dTTP and dUTP. Containing nucleoside triphosphate containing reagents,
Including kit.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US7611108P | 2008-06-26 | 2008-06-26 | |
| US61/076,111 | 2008-06-26 |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JP2010004884A JP2010004884A (en) | 2010-01-14 |
| JP2010004884A5 true JP2010004884A5 (en) | 2012-03-08 |
| JP5596308B2 JP5596308B2 (en) | 2014-09-24 |
Family
ID=41136785
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2009151195A Active JP5596308B2 (en) | 2008-06-26 | 2009-06-25 | Improved method for preventing carryover contamination in nucleic acid amplification technology |
Country Status (7)
| Country | Link |
|---|---|
| US (1) | US8669061B2 (en) |
| EP (1) | EP2138590B1 (en) |
| JP (1) | JP5596308B2 (en) |
| CN (1) | CN101613730B (en) |
| CA (1) | CA2670300C (en) |
| ES (1) | ES2386023T3 (en) |
| HK (1) | HK1138330A1 (en) |
Families Citing this family (19)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8669061B2 (en) | 2008-06-26 | 2014-03-11 | Roche Molecular Systems, Inc. | Method for the prevention of carryover contamination in nucleic acid amplification technologies |
| WO2013022779A1 (en) * | 2011-08-05 | 2013-02-14 | Ibis Biosciences, Inc. | Analysis of genetic biomarkers for forensic analysis and fingerprinting |
| ES2449665B2 (en) * | 2012-09-19 | 2014-07-17 | Servizo Galego De Saúde-Conselleria De Sanidade, Xunta De Galicia | Procedure for the clinical diagnosis of an infectious disease based on the use of quantitative PCR, labeled oligonucleotides of 8 to 9 nucleotides in length and the UNG of the Atlantic Cod (Gadus morhua). |
| US20160257985A1 (en) * | 2013-11-18 | 2016-09-08 | Rubicon Genomics, Inc. | Degradable adaptors for background reduction |
| US10036061B2 (en) * | 2013-12-20 | 2018-07-31 | Roche Molecular Systems, Inc. | Oligonucleotide inhibitor of DNA polymerases |
| EP3152323A1 (en) * | 2014-06-05 | 2017-04-12 | Qiagen GmbH | Optimization of dna amplification reactions |
| EP4026914A1 (en) | 2014-10-08 | 2022-07-13 | Cornell University | Method for identification and relative quantification of nucleic acid methylation changes using combined nuclease, ligation, and polymerase reactions with carryover prevention |
| CN104389026A (en) * | 2014-10-30 | 2015-03-04 | 北京诺禾致源生物信息科技有限公司 | Method for establishing single cell transcriptome sequencing library and application of method |
| JP6623324B2 (en) * | 2015-09-07 | 2019-12-25 | 株式会社ファスマック | Multi-item simultaneous detection method for isothermal amplification reaction products |
| ES2855748T3 (en) | 2016-07-12 | 2021-09-24 | Hoffmann La Roche | Primer extension target enrichment |
| WO2018015318A1 (en) | 2016-07-18 | 2018-01-25 | F. Hoffmann-La Roche Ag | Method for generating single-stranded circular dna libraries for single molecule sequencing |
| JP2021506314A (en) | 2017-12-21 | 2021-02-22 | エフ．ホフマン−ラ ロシュ アーゲーＦ． Ｈｏｆｆｍａｎｎ−Ｌａ Ｒｏｃｈｅ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ | Target enrichment by unidirectional dual probe primer extension |
| WO2020224611A1 (en) * | 2019-05-07 | 2020-11-12 | Institute Of Genetics And Developmental Biology, Chinese Academy Of Sciences | Improved gene editing system |
| EP4031679A1 (en) | 2019-09-20 | 2022-07-27 | F. Hoffmann-La Roche AG | Immune repertoire profiling by primer extension target enrichment |
| EP4162083A1 (en) | 2020-06-08 | 2023-04-12 | F. Hoffmann-La Roche AG | Methods and compositions for detecting structural rearrangements in a genome |
| CN113186037B (en) * | 2021-04-23 | 2022-12-13 | 山东博弘基因科技有限公司 | Nucleic acid scavenger |
| CN114214395A (en) * | 2021-12-31 | 2022-03-22 | 上海星耀医学科技发展有限公司 | Nucleic acid amplification method and kit for improving gene detection accuracy |
| WO2024013241A1 (en) | 2022-07-14 | 2024-01-18 | F. Hoffmann-La Roche Ag | Variant allele enrichment by unidirectional dual probe primer extension |
| CN116773291B (en) * | 2023-03-31 | 2024-04-19 | 艾普拜生物科技(苏州)有限公司 | Paraffin slice DNA pretreatment reagent and method |
Family Cites Families (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH089997B2 (en) | 1987-05-26 | 1996-01-31 | 日立冷熱株式会社 | Multi-blade blower |
| US5683896A (en) | 1989-06-01 | 1997-11-04 | Life Technologies, Inc. | Process for controlling contamination of nucleic acid amplification reactions |
| DE69130800T2 (en) * | 1990-07-24 | 1999-09-16 | Hoffmann La Roche | REDUCING NON-SPECIFIC AMPLIFICATION DURING AN (IN VITRO) NUCLEIC ACID AMPLIFICATION USING MODIFIED NUCLEIC ACID BASES |
| CA2073298C (en) * | 1991-07-12 | 2007-04-17 | James L. Hartley | Process for controlling contamination of nucleic acid amplification reactions |
| JP3416981B2 (en) | 1993-03-30 | 2003-06-16 | 株式会社島津製作所 | Nucleic acid synthesis method |
| JPH089997A (en) | 1994-06-28 | 1996-01-16 | Shimadzu Corp | Method for synthesizing nucleic acid and reagent kit used therefor |
| US6413747B1 (en) | 1994-10-03 | 2002-07-02 | Shimadzu Corporation | Enhancement of nucleic acid amplification by the addition of a polyamine |
| US6190865B1 (en) * | 1995-09-27 | 2001-02-20 | Epicentre Technologies Corporation | Method for characterizing nucleic acid molecules |
| DE19545320A1 (en) | 1995-12-05 | 1997-06-12 | Boehringer Mannheim Gmbh | Thermolabile uracil DNA glycosylase, process for its preparation and use for removing uracil from DNA |
| GB9600384D0 (en) | 1996-01-09 | 1996-03-13 | Nyfotek As | Dna glycosylases |
| JP4629167B2 (en) | 1997-10-17 | 2011-02-09 | 株式会社島津製作所 | Nucleic acid synthesis method |
| EP1038974A1 (en) | 1999-03-26 | 2000-09-27 | Mira Diagnostica GmbH | Method, oligonucleotides suitable for the degradation of in vitro synthezised nucleic acid molecules |
| EP1041159A3 (en) | 1999-03-26 | 2000-10-25 | MIRA Diagnostika GmbH | Method, oligonucleotides suitable for the degradation of in vitro synthesized nucleic acid molecules |
| JP2001008680A (en) | 1999-06-30 | 2001-01-16 | Shimadzu Corp | Synthesis of nucleic acid |
| WO2002090536A2 (en) | 2001-05-10 | 2002-11-14 | Novozymes A/S | A method for producing recombined polynucleotides |
| WO2008005459A2 (en) * | 2006-06-30 | 2008-01-10 | Nugen Technologies, Inc. | Methods for fragmentation and labeling of nucleic acids |
| US8669061B2 (en) | 2008-06-26 | 2014-03-11 | Roche Molecular Systems, Inc. | Method for the prevention of carryover contamination in nucleic acid amplification technologies |
-
2009
- 2009-06-16 US US12/485,569 patent/US8669061B2/en active Active
- 2009-06-24 ES ES09008229T patent/ES2386023T3/en active Active
- 2009-06-24 EP EP09008229A patent/EP2138590B1/en active Active
- 2009-06-25 JP JP2009151195A patent/JP5596308B2/en active Active
- 2009-06-25 CA CA2670300A patent/CA2670300C/en not_active Expired - Fee Related
- 2009-06-26 CN CN200910146289.7A patent/CN101613730B/en active Active
-
2010
- 2010-04-21 HK HK10103924.3A patent/HK1138330A1/en not_active IP Right Cessation
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